Multi-Functional Hemocompatible Porous Polymer Bead Sorbent For Removing Protein Based Toxins And Potassium From Biological Fluids

ABSTRACT

The invention concerns biocompatible polymer systems comprising at least one polymer with a plurality of pores, said polymer comprising a sulfonic acid salt functionality designed to adsorb a broad range of protein based toxins from less than 0.5 kDa to 1,000 kDa and positively charged ions including but not limited to potassium.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation application of U.S. patentapplication Ser. No. 15/769,361 filed Apr. 19, 2018 which claimspriority to International Patent Application No. PCT/US2016/058019 filedOct. 21, 2016 which claims the benefit of U.S. Provisional PatentApplication No. 62/245,071 filed on Oct. 22, 2015. The contents of whichare hereby incorporated by reference in their entireties.

GOVERNMENT RIGHTS

The subject matter disclosed herein was made with government supportunder contract number HHSN268201600006C, awarded by The National Heart,Lung, and Blood Institute (NHLBI). The subject matter disclosed hereinwas also made with government support under contract numberW81XWH-12-C-0038, awarded by The Department of Defense Small BusinessInnovation Research (DOD-SBIR). The government has certain rights in theherein disclosed subject matter.

TECHNICAL FIELD

The disclosed inventions are in the field of porous polymeric sorbents.The disclosed inventions are also in the field of broadly reducingcontaminants in blood and blood products that can cause transfusionreactions; including, but not limited to, potassium, free hemoglobin,cytokines, bioactive lipids, and immunoglobulins. Additionally, thedisclosed inventions are in the field of broadly removing contaminantsby perfusion or hemoperfusion after tissue destruction; including, butnot limited to, potassium, free hemoglobin, free myoglobin, cytokines,bioactive lipids, and immunoglobulins.

BACKGROUND

Packed red blood cell (pRBC) units contain reactive donor antibodies,free hemoglobin, high extracellular potassium levels, and biologicallyactive inflammatory mediators that have the potential to cause adverseeffects during blood transfusions. Such adverse effects can includenon-hemolytic febrile and allergic transfusion reactions, atypicalinfections, allo-immunization, and potentially fatal reactions, liketransfusion related acute lung injury (TRALI). Furthermore, transfusionrisk increases in patients receiving multiple pRBCs, such as thoseinvolved in trauma or undergoing surgery, and in primed susceptiblepatients, such as those in critical care or undergoing high-risksurgery.

The likelihood of adverse effects increases over time for stored bloodor blood products, as concentrations of many biological responsemodifiers, such as potassium, free hemoglobin, and cytokines, increasewith storage duration. Cytokines are produced by residual leukocytesduring storage of platelets and pRBCs, and can cause inflammation,fever, and direct vascular and organ injury. Erythrocytes containphosphatidyl choline, and cytosolic and membrane phospholipase A2,contributing increasing levels of lysophosphatidylcholine (lysoPC)during storage. Structural and biochemical changes that RBCs undergo aredescribed as “storage lesion” and lead to a progressive loss ofhemoglobin, and potassium. Plasma free hemoglobin can rapidly overwhelmthe scavenging capability of haptoglobin, resulting in oxidative damageto lipids, proteins, endothelial cells, tissues, and renal proximaltubules, and in depletion of nitric oxide (NO) upon transfusion.Increases in extracellular potassium during storage lead to an increasedrisk of hyperkalemia and arrhythmia, particularly for large volume or“massive” transfusions and transfusions in newborns and infants.

Hyperkalemia describes a condition in which the potassium level in theblood exceeds a concentration of 5 mEq/L, where concentrations exceeding7 mEq/L are considered severe cases. The electrical rhythm of the heartcan be altered by moderate hyperkalemia, while severe conditions maycause the heart to stop beating. In addition to blood transfusions,another major cause of hyperkalemia is tissue destruction that causesdying cells to release potassium into blood circulation. Tissuedestruction typically results from trauma, burns, hemolysis, massivelysis of tumor cells, rhabdomyolysis, or major surgery, such as cardiacsurgery or cardiopulmonary bypass (CPB), where severe tissue destructionleads to more severe cases of hyperkalemia. In addition to the releaseof potassium into blood circulation, massive tissue injury ischaracterized by release of a large amount of myoglobin from damagedmuscle tissue, plasma free hemoglobin from hemolyzed red blood cells,damage associated molecular pattern (DAMP) factors from damaged cells,and an upregulation of pro- and anti-inflammatory mediators, such ascytokines. Excessive free myoglobin, free hemoglobin, and otherinflammatory mediators, can lead to complications such as renal failureor even death. Abnormal regulation of cytokines, or release of DAMPS,may lead to systemic inflammatory response syndrome (SIRS) andmulti-organ dysfunction (MODs).

Currently, there are existing technologies for potassium removal, orantibody removal, from stored blood or blood products. KawasumiLaboratories has developed a single-pass in-line potassium adsorptionfilter to reduce the risk of hyperkalemia and improve safety for bloodtransfusions. The filter functions by exchanging potassium ions (K⁺) forsodium ions (Na⁺) to decrease the concentration of K⁺ in stored RBCunits. In an in-vitro study conducted by Yamada et. al, 10 filters weretested using each of three AS-3 RBC units via gravity filtration. Themean decrease in potassium was 97.5%, 91.2%, and 64.4% for the first,second, and third units, respectively. Accompanying the decrease inpotassium were mean increases of sodium by 33%, magnesium by 151.4%, andtotal calcium by 116.1%. Plasma hemoglobin was unchanged afterfiltration.

A journal article published by Terai et. al., titled “Development of aPotassium-Specific Adsorbent for Direct Hemoperfusion”, describes astudy assessing the development of a sodium/calcium/magnesium exchangeresin mixture that removes potassium without associated electrolyteabnormalities. At the time the article was written, direct hemoperfusionover an exchange resin was capable of lowering elevated serum potassiumlevels, but had not been used clinically due to subsequent electrolyteabnormalities. Prior to evaluating the exchange resin in an in vivomodel, batch experiments were conducted in vitro to identify aneffective ratio of sodium to calcium to magnesium for the resin mixture.Results from the study demonstrated a reduction of elevated plasmapotassium levels from about 6.7 to about 3.5 mEq/L in anephric dogs,without any significant change in levels of sodium, calcium, magnesium,albumin, total protein, or cholesterol, after 2 hours of directhemoperfusion through an exchange resin column. Pre- andpost-hemoperfusion platelet counts and plasma free hemoglobin levelswere also measured, where post-hemoperfusion platelet counts were onlyabout 45% of pre-hemoperfusion levels, and there was no significantchange in plasma free hemoglobin levels.

Patent WO 2012118735 A2, entitled “Removal of immunoglobulins andleukocytes from biological fluids,” discloses devices, systems, andmethods, for depleting biological fluids of immunoglobulins andleukocytes. It describes a system comprising immunoglobulin bindingmedia and a leukocyte depletion filter element, where the binding mediaconsist of cellulose beads and are placed into the pre-filtration bloodbag. In one example, 30 g dry weight cellulose beads, (4-MEP) HyperCel™chromatography sorbent (Pall Corporation), were placed in a blood bag towhich a unit of 5 day old AS-3 RBC was added, and the blood bag mixed ona rotamixer. The RBCs were gravity filtered through a downstream filter,where beads were trapped in an immunoglobulin binding media chamber andfiltered cells passed through a fibrous leukocyte depletion filterbefore being collected and analyzed. Leukocyte content was reduced by5.17 log, IgA reduced by 81%, IgG by 98%, and IgM by 42%. In anotherexample, the ability of the leukocyte filter to remove cytokines wasexamined. Two units of 22-30 day old ABO compatible red cell concentratewere pooled together and then split into two lots. The first was placedin a blood bag containing about 25-33 g dry weight cellulose beads,(4-MEP) HyperCel™ chromatography sorbent (Pall Corporation), with 10 mLPBS and mixed for 45 minutes, and the second passed through a BPF4 HighEfficiency leukocyte depletion filter (Pall Corporation) via gravityfiltration. Afterwards, both lots were analyzed and it was found that inthe aliquot placed in contact with the beads, interleukin 1-Beta (IL-1β)was reduced by 45.7%, interleukin-6 (IL-6) by 26.9%, interleukin-8(IL-8) by 57.1% and tissue necrosis factor-alpha (TNF-α) by 49.9% Forthe aliquot passed through the filter, IL-1β was not reduced, IL-6 wasnot reduced, IL-8 was reduced by 35.0% and TNF-α reduced by 7.5%

In a journal article by Silliman et. al., it was demonstrated thatpre-storage filtration of packed RBCs removes HLA and HNA antibodies,reducing pro-inflammatory activity in RBC supernatant in an animal TRALImodel. In the described study, plasma that contained antibodies to humanlymphocyte antigen (HLA)-A2, or human neutrophil antigen (HNA)-3a, wasfiltered and priming activities of specific HNA-3a and HLA-2a weremeasured. OX27 antibodies were added to plasma and filtration wasanalyzed using a 2-event animal model for TRALI. RBC units from 31donors, who were known to possess antibodies against HLA antigens, werefiltered. In addition, 4 RBC units underwent standard leukoreduction.PMN priming activity, immunoglobulins, HLA antibodies, and ability toinduce TRALI were measured. Filtration of the plasma was shown to removemore than 96% of IgG, and antibodies to HLA-A2 and HNA-3a, includingtheir respective priming activity, and mitigated in vivo TRALI.Antibodies to HLA antigens were removed in experimental filtration ofRBC units, accompanied by an inhibition of accumulation of lipid primingactivity and lipid-mediated TRALI.

The sorbent material described herein is uniquely designed toefficiently remove free hemoglobin, antibodies, bioactive lipids,cytokines, and potassium, from blood and blood products. The polymer ismulti-functional, retaining said biomolecules through tortuous path,sorption, pore capture, and ion exchange mechanisms. Novel chemistry isused to synthesize the polymer, utilizing a controlled sulfonationprocedure that allows for the incorporation of sulfonic acid groups ontothe aromatic rings without oxidizing all residual double bonds. Thisallows the polymeric matrix to maintain protein sorption and ionexchange capabilities, while still leaving residual functional groupsavailable for hemocompatibility improvement modifications. The balancebetween sulfonation and retention of residual double bonds is crucialfor preparation of an effective polymer sorbent.

Differentiating the multi-functional polymer from other filters thatremove only reactive proteins or only potassium is its ability to removeboth simultaneously without sacrificing binding capacity for either.Additionally, the sorbent is able to remove cytokines and inflammatoryprotein moieties simultaneously while removing potassium and antibodies.For hemoperfusion applications, it is a requirement that the polymer ishemocompatible. Using the unactivated partial thromboplastin time (uPTT)assay as a measure of thrombogenicity, the polymer described hereinexhibits minimal activation, indicating a plasma-like interaction. Thispolymer is suited for a wide variety of applications, as many cases oftrauma, burn, and major surgery, result in hyperkalemia, cytokine storm,and require blood transfusions. The ability to use one multi-applicationfilter has many advantages over using many single-application filters.Given the value of blood and blood products, the use of a single,smaller filter that minimizes cell loss within the retained volume andreduces complexity of material quality assurance is very desirable.

SUMMARY

In some aspects, the invention concerns biocompatible polymer systemcomprising at least one polymer, said polymer comprising (i) a pluralityof pores and (ii) a sulfonic acid salt functionality; the polymer systemcapable of adsorbing (i) a broad range of protein based toxins having amolecular weight of from less than about 0.5 kDa to about 1,000 kDa (orabout 1 kDa to about 1,000 kDa in some embodiments) and (ii) positivelycharged ions. Some polymer systems have a polymer pore structure thathas a total volume of pore sizes in the range of from 10 Å to 40,000 Ågreater than 0.1 cc/g and less than 5.0 cc/g dry polymer. Some preferredpolymers are hemocompatible. The polymer system has the form of a solidsupport. Certain preferred polymer systems have a geometry of aspherical bead. Other polymer systems have the form of a fiber,monolithic column, film, membrane, or semi-permeable membrane.

In some embodiments, the toxins adsorbed comprise one or more ofinflammatory mediators and stimulators comprised of one or more ofcytokines, superantigens, monokines, chemokines, interferons, proteases,enzymes, peptides including bradykinin, soluble CD40 ligand, bioactivelipids, oxidized lipids, cell-free hemoglobin, cell-free myoglobin,growth factors, glycoproteins, prions, toxins, bacterial and viraltoxins, endotoxins, drugs, vasoactive substances, foreign antigens,antibodies, and positively charged ions. In some preferred embodiments,the positively charged ion is potassium.

The polymers can be made by any means known in the art to produce asuitable porous polymer. In some embodiments, the polymer is made usingsuspension polymerization. Some polymers comprise a hypercrosslinkedpolymer. Certain spherical beads have a biocompatible hydrogel coating.In certain embodiments, the polymer is in the form of hypercrosslinkedor a macroreticular porous polymer beads that have been sulfonated undermild conditions that retain residual functionality of any unreacteddouble bonds and chloromethyl groups. The unreacted double bonds orchloromethyl groups can be modified via free radical or S_(N)2 typechemistry to attach one or more of biocompatible and hemocompatiblemonomers, cross-linkers or low molecular weight oligomers.

In some embodiments, the porous polymer beads comprise sulfonic acidgroups or a salt thereof, sulfonyl chloride, or sulfonate ester groups.The polymer beads comprising sulfonic acid groups or a salt thereof,sulfonyl chloride, or sulfonate ester groups can be produced by graftcopolymerization of (i) premade porous polymer that contains unreacteddouble bonds with (ii) polymerizable vinyl monomers containing sulfonicacid groups or a salt thereof to form a mixture comprisinghemocompatible vinyl monomers.

Some polymer systems are constructed from polymerizable vinyl monomerscontaining sulfonic acid groups or a salt thereof which arecopolymerized in the presence of cross-linker, hemocompatible monomer,monomer and suitable porogen to yield porous polymeric polymercontaining a sulfonic acid salt functionality.

Certain polymers are formed and subsequently modified to bebiocompatible. Some modifications comprise forming a biocompatiblesurface coating or layer.

Other aspects include methods of perfusion comprising passing aphysiologic fluid once through or by way of a suitable extracorporealcircuit through a device comprising the biocompatible polymer systemdescribed herein.

Yet another aspect concerns devices for removing (i) a broad range ofprotein based toxins from less than 0.5 kDa to 1,000 kDa and (ii)positively charged ions from physiologic fluid comprising thebiocompatible polymer system described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are included to provide a furtherunderstanding of the disclosure, are incorporated in and constitute apart of this specification, illustrate aspects of the disclosure andtogether with the detailed description serve to explain the principlesof the disclosure. No attempt is made to show structural details of thedisclosure in more detail than may be necessary for a fundamentalunderstanding of the disclosure and the various ways in which it may bepracticed. In the drawings:

FIGS. 1, 2 and 3 present log differential pore volume plots for CY15100and CY15102.

FIGS. 4, 5 and 6 show plots of log differential pore volume for modifiedpolymers.

FIGS. 7 and 8 show plots of log differential pore volume for polymersCY15048 and CY15049.

FIG. 9 presents the percentage of initial free hemoglobin removed duringsingle-pass filtration, averaged from three trials

FIG. 10 displays pre- and post-filtration potassium ion concentration inblood, averaged from three trials.

FIG. 11 presents dynamic recirculation data for CY14144, averaged from 7trials.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

As required, detailed embodiments of the present invention are disclosedherein; it is to be understood that the disclosed embodiments are merelyexemplary of the invention that may be embodied in various forms.Therefore, specific structural and functional details disclosed hereinare not to be interpreted as limits, but merely as a basis for teachingone skilled in the art to employ the present invention. The specificexamples below will enable the invention to be better understood.However, they are given merely by way of guidance and do not imply anylimitation.

The present invention may be understood more readily by reference to thefollowing detailed description taken in connection with the accompanyingfigures and examples, which form a part of this disclosure. It is to beunderstood that this invention is not limited to the specific materials,devices, methods, applications, conditions or parameters describedand/or shown herein, and that the terminology used herein is for thepurpose of describing particular embodiments by way of example only andis not intended to be limiting of the claimed invention. The term“plurality”, as used herein, means more than one. When a range of valuesis expressed, another embodiment includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it willbe understood that the particular value forms another embodiment. Allranges are inclusive and combinable.

It is to be appreciated that certain features of the invention whichare, for clarity, described herein in the context of separateembodiments, may also be provided in combination in a single embodiment.Conversely, various features of the invention that are, for brevity,described in the context of a single embodiment, may also be providedseparately or in any subcombination. Further reference to values statedin ranges includes each and every value within that range.

The following definitions are intended to assist in understanding thepresent invention:

The term “biocompatible” is defined to mean the sorbent is capable ofcoming in contact with physiologic fluids, living tissues, or organisms,without producing unacceptable clinical changes during the time that thesorbent is in contact with the physiologic fluids, living tissues, ororganisms.

The term “hemocompatible” is defined as a condition whereby abiocompatible material when placed in contact with whole blood or bloodplasma results in clinically acceptable physiologic changes.

As used herein, the term “physiologic fluids” are liquids that originatefrom the body and can include, but are not limited to, nasopharyngeal,oral, esophageal, gastric, pancreatic, hepatic, pleural, pericardial,peritoneal, intestinal, prostatic, seminal, vaginal secretions, as wellas tears, saliva, lung, or bronchial secretions, mucus, bile, blood,lymph, plasma, serum, synovial fluid, cerebrospinal fluid, urine, andinterstitial, intracellular, and extracellular fluid, such as fluid thatexudes from burns or wounds.

As used herein, the term “laboratory or manufacturing fluids” aredefined as liquids that are used in life sciences applications thatinclude, but are not limited to, tissue and cell culture media andadditives, chemical or biologic assay media, sample preparation buffers,biologic manufacturing media, growth media, and bioreactor media.

As used herein, the term “sorbent” includes adsorbents and absorbents.

For purposes of this invention, the term “sorb” is defined as “taking upand binding by absorption and adsorption”.

For the purposes of this invention, the term “perfusion” is defined aspassing a physiologic fluid, once through or by way of a suitableextracorporeal circuit, through a device containing the porous polymericadsorbent to remove toxic molecules from the fluid.

The term “hemoperfusion” is a special case of perfusion where thephysiologic fluid is blood.

The term “dispersant” or “dispersing agent” is defined as a substancethat imparts a stabilizing effect upon a finely divided array ofimmiscible liquid droplets suspended in a fluidizing medium.

The term “heparin mimicking polymer” refers to any polymer thatpossesses the same anticoagulant and/or antithrombogenic properties asheparin.

The term “macroreticular synthesis” is defined as a polymerization ofmonomers into polymer in the presence of an inert precipitant whichforces the growing polymer molecules out of the monomer liquid at acertain molecular size dictated by the phase equilibria to give solidnanosized microgel particles of spherical or almost spherical symmetrypacked together to give a bead with physical pores of an open cellstructure [U.S. Pat. No. 4,297,220, Meitzner and Oline, Oct. 27, 1981;R. L. Albright, Reactive Polymers, 4, 155-174(1986)].

The term “hypercrosslinked” describes a polymer in which the singlerepeating unit has a connectivity of more than two. Hypercrosslinkedpolymers are prepared by crosslinking swollen, or dissolved, polymerchains with a large number of rigid bridging spacers, rather thancopolymerization of monomers. Crosslinking agents may includebis(chloromethyl) derivatives of aromatic hydrocarbons, methylal,monochlorodimethyl ether, and other bifunctional compounds that reactwith the polymer in the presence of Friedel-Crafts catalysts [Tsyurupa,M. P., Z. K. Blinnikova, N. A. Proskurina, A. V. Pastukhov, L. A.Pavlova, and V. A. Davankov. “Hypercrosslinked Polystyrene: The FirstNanoporous Polymeric Material.” Nanotechnologies in Russia 4 (2009):665-75.]

Some preferred polymers comprise residues from one or more monomers, orcontaining monomers, or mixtures thereof, selected from acrylonitrile,allyl ether, allyl glycidyl ether, butyl acrylate, butyl methacrylate,cetyl acrylate, cetyl methacrylate, 3,4-dihydroxy-1-butene,dipentaerythritol diacrylate, dipentaerythritol dimethacrylate,dipentaerythritol tetraacrylate, dipentaerythritol tetramethacrylate,dipentaerythritol triacrylate, dipentaerythritol trimethacrylate,divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone,3,4-epoxy-1-butene, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, ethylacrylate, ethyl methacrylate, ethylstyrene, ethylvinylbezene, glycidylmethacrylate, methyl acrylate, methyl methacrylate, octyl acrylate,octyl methacrylate, pentaerythritol diacrylate, pentaerythritoldimethacrylate, pentaerythritol tetraacrylate, pentaerythritoltetramethacrylate, pentaerythritol triacrylate, pentaerythritoltrimethacrylate, styrene, trimethylolpropane diacrylate,trimethylolpropane dimethacrylate, trimethylolpropane triacrylate,trimethylolpropane trimethacrylate, trivinylbenzene,trivinylcyclohexane, vinyl acetate, vinylbenzyl alcohol,4-vinyl-1-cyclohexene 1,2-epoxide, vinylformamide, vinylnaphthalene,2-vinyloxirane, and vinyltoluene.

Some embodiments of the invention use an organic solvent and/orpolymeric porogen as the porogen or pore-former, and the resulting phaseseparation induced during polymerization yield porous polymers. Somepreferred porogens are selected from, or mixtures comprised of anycombination of, benzyl alcohol, cyclohexane, cyclohexanol,cyclohexanone, decane, dibutyl phthalate, di-2-ethylhexyl phthalate,di-2-ethylhexylphosphoric acid, ethylacetate, 2-ethyl-1-hexanoic acid,2-ethyl-1-hexanol, n-heptane, n-hexane, isoamyl acetate, isoamylalcohol, n-octane, pentanol, poly(propylene glycol), polystyrene,poly(styrene-co-methyl methacrylate), tetraline, toluene,tri-n-butylphosphate, 1,2,3-trichloropropane, 2,2,4-trimethylpentane,xylene.

In yet another embodiment, the dispersing agent is selected from a groupconsisting of hydroxyethyl cellulose, hydroxypropyl cellulose,poly(diethylaminoethyl acrylate), poly(diethylaminoethyl methacrylate),poly(dimethylaminoethyl acrylate), poly(dimethylaminoethylmethacrylate), poly(hydroxyethyl acrylate), poly(hydroxyethylmethacrylate), poly(hydroxypropyl acrylate), poly(hydroxypropylmethacrylate), poly(vinyl alcohol), salts of poly(acrylic acid), saltsof poly(methacrylic acid) and mixtures thereof.

Preferred sorbents are biocompatible. In another further embodiment, thepolymer is biocompatible. In yet another embodiment, the polymer ishemocompatible. In still a further embodiment, the biocompatible polymeris hemocompatible. In still a further embodiment, the geometry of thepolymer is a spherical bead.

In another embodiment, the biocompatible polymer comprisespoly(N-vinylpyrrolidone).

The coating/dispersant on the porous poly(styrene-co-divinylbenzene)resin will imbue the material with improved biocompatibility.

In still yet another embodiment, a group of cross-linkers consisting ofdipentaerythritol diacrylates, dipentaerythritol dimethacrylates,dipentaerythritol tetraacrylates, dipentaerythritol tetramethacrylates,dipentaerythritol triacrylates, dipentaerythritol trimethacrylates,divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone,pentaerythritol diacrylates, pentaerythritol dimethacrylates,pentaerythritol tetraacrylates, pentaerythritol tetramethacrylates,pentaerythritol triacrylates, pentaerythritol trimethacrylates,trimethylolpropane diacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane trimethacrylate,trivinylbenzene, trivinylcyclohexane and mixtures thereof can be used information of a hemocompatible hydrogel coating.

In some embodiments, the polymer is a polymer comprising at least onecrosslinking agent and at least one dispersing agent. The dispersingagent may be biocompatible. The dispersing agents can be selected fromchemicals, compounds or materials such as hydroxyethyl cellulose,hydroxypropyl cellulose, poly(diethylaminoethyl acrylate),poly(diethylaminoethyl methacrylate), poly(dimethylaminoethyl acrylate),poly(dimethylaminoethyl methacrylate), poly(hydroxyethyl acrylate),poly(hydroxyethyl methacrylate), poly(hydroxypropyl acrylate),poly(hydroxypropyl methacrylate), poly(vinyl alcohol), salts ofpoly(acrylic acid), salts of poly(methacrylic acid) and mixturesthereof; the crosslinking agent selected from a group consisting ofdipentaerythritol diacrylates, dipentaerythritol dimethacrylates,dipentaerythritol tetraacrylates, dipentaerythritol tetramethacrylates,dipentaerythritol triacrylates, dipentaerythritol trimethacrylates,divinylbenzene, divinylformamide, divinylnaphthalene, divinylsulfone,pentaerythritol diacrylates, pentaerythritol dimethacrylates,pentaerythritol tetraacrylates, pentaerythritol tetramethacrylates,pentaerythritol triacrylates, pentaerythritol trimethacrylates,trimethylolpropane diacrylate, trimethylolpropane dimethacrylate,trimethylolpropane triacrylate, trimethylolpropane trimethacrylate,trivinylbenzene, trivinylcyclohexane and mixtures thereof. Preferably,the polymer is developed simultaneously with the formation of thecoating, wherein the dispersing agent is chemically bound or entangledon the surface of the polymer.

In still another embodiment, the biocompatible polymer coating isselected from a group consisting of poly(hydroxyethyl methacrylate),poly(hydroxyethyl acrylate), poly(dimethylaminoethyl methacrylate),salts of poly(acrylic acid), salts of poly(methacrylic acid),poly(diethylaminoethyl methacrylate), poly(hydroxypropyl methacrylate),poly(hydroxypropyl acrylate), poly(N-vinylpyrrolidone), poly(vinylalcohol) and mixtures thereof. In another embodiment, the salts may besodium and potassium salts and in still another embodiment, the saltsare water-soluble salts.

In still another embodiment, the biocompatible oligomer coating isselected from a group consisting of poly(hydroxyethyl methacrylate),poly(hydroxyethyl acrylate), poly(dimethylaminoethyl methacrylate),salts of poly(acrylic acid), salts of poly(methacrylic acid),poly(diethylaminoethyl methacrylate), poly(hydroxypropyl methacrylate),poly(hydroxypropyl acrylate), poly(N-vinylpyrrolidone), poly(vinylalcohol) and mixtures thereof. In another embodiment, the salts may besodium and potassium salts and in still another embodiment, the saltsare water-soluble salts.

The present biocompatible sorbent compositions are comprised of aplurality of pores. The biocompatible sorbents are designed to adsorb abroad range of toxins from less than 0.5 kDa to 1,000 kDa. While notintending to be bound by theory, it is believed the sorbent acts bysequestering molecules of a predetermined molecular weight within thepores. The size of a molecule that can be sorbed by the polymer willincrease as the pore size of the polymer increases. Conversely, as thepore size is increased beyond the optimum pore size for adsorption of agiven molecule, adsorption of said protein may or will decrease.

In certain methods, the solid form is porous. Some solid forms arecharacterized as having a pore structure having a total volume of poresizes in the range of from 10 Å to 40,000 Å greater than 0.1 cc/g andless than 5.0 cc/g dry polymer.

In certain embodiments, the polymers can be made in bead form having adiameter in the range of 0.1 micrometers to 2 centimeters. Certainpolymers are in the form of powder, beads or other regular orirregularly shaped particulates.

In some embodiments, the plurality of solid forms comprises particleshaving a diameter in the range for 0.1 micrometers to 2 centimeters.

In some methods, the undesirable molecules are inflammatory mediatorsand stimulators comprised of cytokines, superantigens, monokines,chemokines, interferons, proteases, enzymes, peptides includingbradykinin, soluble CD40 ligand, bioactive lipids, oxidized lipids,cell-free hemoglobin, damage-associated molecular pattern (DAMPs),Pathogen-associated molecular pattern molecules (PAMPs), cell-freemyoglobin, growth factors, glycoproteins, prions, toxins, bacterial andviral toxins, endotoxins, drugs, vasoactive substances, foreignantigens, antibodies, and positively charged ions, including, but notlimited to, potassium.

In some methods, the antibodies can be immunoglobulin A (IgA),immunoglobulin D (IgD), immunoglobulin E (IgE), immunoglobulin D (IgG),immunoglobulin D (IgM) and/or immunoglobulin fragments or subunits.

DAMPs have been associated with countless syndromes and diseases. Theseinclude complications from trauma, burns, traumatic brain injury andinvasive surgery, and also organ-specific illnesses like liver disease,kidney dialysis complications, and autoimmune diseases. DAMPs are hostmolecules that can initiate and perpetuate noninfectious SIRS andexacerbate infectious SIRS. DAMPs are a diverse family of molecules thatare intracellular in physiological conditions and many are nuclear orcytosolic proteins. DAMPS can be divided into two groups: (1) moleculesthat perform noninflammatory functions in living cells (such as HMGB1)and acquire immunomodulatory properties when released, secreted,modified, or exposed on the cell surface during cellular stress, damage,or injury, or (2) alarmins, i.e., molecules that possess cytokine-likefunctions (such as β-Defensins and Cathelicidin), which can be stored incells and released upon cell lysis, whereupon they contribute to theinflammatory response. When released outside the cell or exposed on thesurface of the cell following tissue injury, they move from a reducingto an oxidizing milieu, which affects their activity. Also, followingnecrosis, mitochondrial and nuclear DNA fragments are released outsidethe cell becoming DAMPs.

DAMPs, such as HMGB-1, heat-shock and 5100 proteins are normally foundinside cells and are released by tissue damage. DAMPs act as endogenousdanger signals to promote and exacerbate the inflammatory response.HMGB-1 is a non-histone nuclear protein that is released under stressconditions. Extracellular HMGB-1 is an indicator of tissue necrosis andhas been associated with an increased risk of sepsis and multiple organdysfunction syndrome (MODS). S100 A8 (granulin A, MRP8) and A9 (granulinB\, MRP14) homo and heterodimers bind to and signal directly via theTLR4/lipopolysaccharide receptor complex where they become dangersignals that activate immune cells and vascular endothelium.Procalcitonin is a marker of severe sepsis caused by bacteria and itsrelease into circulation is indicative of the degree of sepsis. Serumamyloid A (SAA), an acute-phase protein, is produced predominantly byhepatocytes in response to injury, infection, and inflammation. Duringacute inflammation, serum SAA levels may rise by 1000-fold. SAA ischemotactic for neutrophils and induces the production ofproinflammatory cytokines. Heat shock proteins (HSP) are a family ofproteins that are produced by cells in response to exposure to stressfulconditions and are named according to their molecular weight (10, 20-30,40, 60, 70, 90). The small 8-kilodalton protein ubiquitin, which marksproteins for degradation, also has features of a heat shock protein.Hepatoma-derived growth factor (HDGF), despite its name, is a proteinexpressed by neurons. HDGF can be released actively by neurons via anonclassical pathway and passively by necrotic cells. Other factors,such as complement factors 3 and 5, are activated as part of the hostdefense against pathogens but can also contribute to the adverseoutcomes in sepsis. Excessive, persistent circulating levels ofcytokines and DAMPs contribute to organ injury and identify thosepatients who have the highest risk of multiple organ dysfunction (MODs)and death in community acquired pneumonia and sepsis.

PAMPs include lipopolysaccharides, lipopeptides, lipoteichoic acid,peptidoglycans, nucleic acids such as double-stranded RNA, toxins andflagellins nd can trigger an immune response in the host (e.g. theinnate immune system) to fight the infection, leading to the productionof high levels of inflammatory and anti-inflammatory mediators, such ascytokines. PAMPs and high cytokine levels, as well as direct tissueinjury (trauma, burns, etc.), can damage tissue, causing theextracellular release of damage-associated molecular pattern (DAMPs)molecules into the bloodstream. DAMPs are a broad class of endogenousmolecules, which like PAMPs, trigger the immune response through patternrecognition receptors (PRRs) such as Toll-like receptors (TLRs).

Preferred sorbents include cross-linked polymeric material derived fromthe reaction of a cross-linker with one or more of the followingpolymerizable monomers, then subsequently sulfonated to form a sulfonicacid salt: acrylonitrile, butyl acrylate, butyl methacrylate, cetylacrylate, cetyl methacrylate, divinylbenzene, ethyl acrylate, ethylmethacrylate, ethylstyrene, methyl acrylate, methyl methacrylate, octylacrylate, octyl methacrylate, styrene, vinylbenzyl alcohol,vinylformamide, vinylnaphthalene, or vinyltoluene.

In some embodiments, radically polymerizable vinyl monomers containing˜SO₃Na groups, or ˜SO₃H groups, can be used in graft copolymerizationwith porous polymers containing polymerizable double bonds. Thesemonomers can be selected from 4-styrene sulfonic acid sodium salt, vinylsulfonic acid sodium salt, 2-acrylamido-2-methyl-1-propanesulfonic acid,2-acrylamido-2-methyl-1-propanesulfonic sodium salt, 3-sulfopropylacrylate sodium salt, 3-sulfopropyl methacrylate sodium salt,[2-(methacryloyloxy)ethyl] dimethyl-(3-sulfopropyl)ammonium hydroxide,N-(3-sulfopropyl)-N-(methacryloxyethyl)-N,N-dimethyl ammonium betaine,para-styrene sulfonyl chloride, or any combinations thereof.Furthermore, para-styrene sulfonyl chloride can be polymerized in thepresence of divinylbenzene and hydrolyzed with sodium hydroxide solutionto directly yield poly(styrene-co-divinylbenzene) porous material with˜SO₃Na groups.

In another embodiment, the present invention relates to a sulfonatedpolymer comprised of at least one crosslinking agent for making thepolymer and at least one dispersing agent whereby the dispersing agentforms a biocompatible surface on the polymer.

In one embodiment the porous polymers of this invention are made bysuspension polymerization in a formulated aqueous phase with freeradical initiation in the presence of aqueous phase dispersants that areselected to provide a biocompatible and a hemocompatible exteriorsurface to the formed polymer beads. The sulfonation of the resultantbeads yields an ion exchange resin coated with a hemocompatiblehydrogel. The beads are made porous by the macroreticular synthesis withan appropriately selected porogen (pore forming agent) and anappropriate time-temperature profile for the polymerization in order todevelop the proper pore structure. The subsequent introduction of thesulfonic acid groups in the already formed network forms a sulfonic acidsalt inner core (ion exchange resin) and a hemocompatible outer hydrogelexterior. Suitable choice of the reaction conditions for the sulfonationallows preservation or expression (via a protecting group) of thehemocompatible nature of the exterior hydrogel.

In another embodiment polymers made by suspension polymerization can bemade biocompatible and hemocompatible by further grafting ofbiocompatible and hemocompatible monomers or low molecular weightoligomers. It has been shown that the radical polymerization proceduredoes not consume all the vinyl groups of DVB introduced intocopolymerization. On average, about 30% of DVB species fail to serve ascrosslinking bridges and remain involved in the network by only one oftwo vinyl groups. The presence of a relatively high amount of pendantvinyl groups is therefore a characteristic feature of the macroporousadsorbents. It can be expected that these pendant vinyl groups arepreferably exposed to the surface of the polymer beads and theirmacropores should be readily available to chemical modification. Thechemical modification of the surface of macroporous DVB-copolymersrelies on chemical reactions of the surface-exposed pendant vinyl groupsand aims at converting these groups into more hydrophilic functionalgroups. This conversion via free radical grafting of monomers and/orcross-linkers or low molecular weight oligomers provides the initialhydrophobic adsorbing material with the property of hemocompatibility.The subsequent introduction of the sulfonic acid groups into the alreadyformed network forms a sulfonic acid salt inner core (ion exchangeresin) and a hemocompatible outer hydrogel exterior. Suitable choice ofthe reaction conditions for the sulfonation allows preservation orexpression (via a protecting group) of the hemocompatible nature of theexterior hydrogel.

Still another embodiment consists of binding long hydrophilic polymerchains onto the beads' surfaces, which should preclude contact betweenblood cells and the sulfonated polystyrene surface. This can beaccomplished via free radical or S_(N)2 type chemistry. The chemicalmodification of the surface of sorbent beads, which is the case in theabove modification, is facilitated by the remarkable peculiarity of thehypercrosslinked polystyrene; namely, that the reactive functionalgroups of the polymer are predominantly located on its surface. Thehypercrosslinked polystyrene is generally prepared by crosslinkingpolystyrene chains with large amounts of bifunctional compounds, inparticular, those bearing two reactive chloromethyl groups. The latteralkylate, in a two-step reaction, two phenyl groups of neighboringpolystyrene chains according to Friedel-Crafts reaction, with evolutionof two molecules of HCl and formation of a cross bridge. During thecrosslinking reaction, the three-dimensional network formed acquiresrigidity. This property gradually reduces the rate of the second step ofthe crosslinking reaction, since the reduced mobility of the secondpendant functional group of the initial crosslinking reagent makes itmore and more difficult to add an appropriate second partner for thealkylation reaction. This is especially characteristic of the secondfunctional groups that happen to be exposed to the surface of the bead.Therefore, of the pendant unreacted chloromethyl groups in the finalhypercrosslinked polymer, the largest portion, if not the majority ofthe groups, are located on the surface of the bead (or on the surface oflarge pores). This circumstance makes it possible to predominantlymodify the surface of the polymer beads by involving the abovechloromethyl groups into various chemical reactions that allowattachment of biocompatible and hemocompatible monomers, and/orcross-linkers or low molecular weight oligomers. The subsequentintroduction of the sulfonic acid groups in the already formed networkforms a sulfonic acid salt inner core (ion exchange resin) and ahemocompatible outer hydrogel exterior. Suitable choice of the reactionconditions for the sulfonation allows preservation or expression (via aprotecting group) of the hemocompatible nature of the exterior hydrogel.

In yet another embodiment, the radical polymerization initiator isinitially added to the dispersed organic phase, not the aqueousdispersion medium as is typical in suspension polymerization. Duringpolymerization, many growing polymer chains with their chain-endradicals show up at the phase interface and can initiate thepolymerization in the dispersion medium. Moreover, the radicalinitiator, like benzoyl peroxide, generates radicals relatively slowly.This initiator is only partially consumed during the formation of beadseven after several hours of polymerization. This initiator easily movestoward the surface of the bead and activates the surface exposed pendantvinyl groups of the divinylbenzene moiety of the bead, thus initiatingthe graft: polymerization of other monomers added after the reaction hasproceeded for a period of time. Therefore, free-radical grafting canoccur during the transformation of the monomer droplets into polymerbeads thereby incorporating monomers and/or cross-linkers or lowmolecular weight oligomers that impart biocompatibility orhemocompatibility as a surface coating. The subsequent introduction ofthe sulfonic acid groups in the already formed network forms a sulfonicacid salt inner core (ion exchange resin) and a hemocompatible outerhydrogel exterior. Suitable choice of the reaction conditions for thesulfonation allows preservation or expression (via a protecting group)of the hemocompatible nature of the exterior hydrogel.

In still yet another embodiment, hypercrosslinked or macroreticularporous polymer beads (including commercial versions) that have beensulfonated under mild conditions that retain residual functionality suchas unreacted double bonds or chloromethyl groups can be modified viafree radical or S_(N)2 type chemistry which would allow attachment ofbiocompatible and a hemocompatible monomers, and/or cross-linkers or lowmolecular weight oligomers. Among various “mild” sulfonating agents,Acetyl Sulfate (prepared from 98% conc. Sulfuric acid and aceticanhydride at low temperatures) is known to be very efficient for DVB orStyrene based polymeric materials. Sulfonation is usually done at 50° C.for several hours using equimolar amounts of acetyl sulfate and DVB orstyrene based polymers. Sulfonation occurs mainly at the benzene ringand unreacted double bonds (in DVB based cross-linked polymeric porousbeads) would be preserved for further functionalization. Usually aftersulfonation with acetyl sulfate, the polymer is converted into —SO₃Naform and can be graft copolymerized with N-vinyl pyrrolidone or otherhemocompatible monomers and/or cross-linkers or low molecular weightoligomers (in bulk with benzoyl peroxide as initiator) or in watersolutions (using sodium persulfate initiator). Resulting sulfonatedpolymer is “coated” with poly(N-vinylpyrrolidone), as an example, tocreate a hemocompatible material capable of removing K⁺ cations fromphysiological fluids.

Some embodiments of the invention involve direct synthesis of porouspolymeric beads containing —SO₃Na groups. Any polymerizable vinylmonomer containing —SO₃Na (or —SO₃H) groups can be polymerized in thepresence of cross-linker monomer (like DVB, bis-acrylamide,bis-(meth)acrylates, etc.) and suitable porogen to yield porouspolymeric beads containing above mentioned functionalities (—SO₃Na orSO₃H). Vinyl monomers containing SO₃Na or SO₃H groups can also becopolymerized with hemocompatible monomer (NVP. 2-HEMA, etc.) inpresence of porogen to yield hemocompatible porous beads containing—SO₃Na groups.

Another embodiment of the invention involves making porous polymer beadscontaining SO₃Na groups via graft copolymerization of premade porouspolymers (containing double bonds unreacted) with polymerizable vinylmonomers containing —SO₃Na or —SO₃H groups (with the mixture of suitablehemocompatible vinyl monomers). Such monomers can be vinyl sulfonic acidNa salt, 4-styrene sulfonic acid Na salt, etc.

The hemoperfusion and perfusion devices consist of a packed bead bed ofthe porous polymer beads in a flow-through container fitted with eithera retainer screen at both the exit end and the entrance end to maintainthe bead bed inside the container or with a subsequent retainer screento collect the beads after mixing. The hemoperfusion and perfusionoperations are performed by passing the whole blood, blood plasma orphysiologic fluid through the packed bead bed. During the perfusionthrough the bead bed, the toxic molecules are retained by sorption,torturous path, and/or ion exchange mechanism the while the remainder ofthe fluid and intact cell components pass through essentially unchangedin concentration.

In some other embodiments, an in-line filter is comprised of a packedbead bed of the porous polymer beads in a flow-through container, fittedwith a retainer screen at both the exit end and the entrance end tomaintain the bead bed inside the container. pRBCs are passed from astorage bag once-through the packed bead bed via gravity, during whichthe toxic molecules are retained by sorption, torturous path, and/or ionexchange mechanisms, while the remainder of the fluid and intact cellcomponents pass through essentially unchanged in concentration.

Certain polymers useful in the invention (as is or after furthermodification) are macroporous polymers prepared from the polymerizablemonomers of styrene, divinylbenzene, ethylvinylbenzene, and the acrylateand methacrylate monomers such as those listed below by manufacturer.Rohm and Haas Company, (now part of Dow Chemical Company): macroporouspolymeric sorbents such as Amberlite™ XAD-1, Amberlite™ XAD-2,Amberlite™ XAD-4, Amberlite™ XAD-7, Amberlite™ XAD-7HP, Amberlite™XAD-8, Amberlite™ XAD-16, Amberlite™ XAD-16 HP, Amberlite™ XAD-18,Amberlite™ XAD-200, Amberlite™ XAD-1180, Amberlite™ XAD-2000, Amberlite™XAD-2005, Amberlite™ XAD-2010, Amberlite™ XAD-761, and Amberlite™XE-305, and chromatographic grade sorbents such as Amberchrom™ CG71,s,m,c, Amberchrom™ CG 161,s,m,c, Amberchrom™ CG 300,s,m,c, andAmberchrom™ CG 1000,s,m,c. Dow Chemical Company: Dowex™ Optipore™ L-493,Dowex™ Optipore™ V-493, Dowex™ Optipore™ V-502, Dowex™ Optipore™ L-285,Dowex™ Optipore™ L-323, and Dowex™ Optipore™ V-503. Lanxess (formerlyBayer and Sybron): Lewatit™ VPOC 1064 MD PH, Lewatit™ VPOC 1163,Lewatit™ OC EP 63, Lewatit™ S 6328A, Lewatit™ OC 1066, and Lewatit™60/150 MIBK. Mitsubishi Chemical Corporation: Diaion™ HP 10, Diaion™ HP20, Diaion™ HP 21, Diaion™ HP 30, Diaion™ HP 40, Diaion™ HP 50, Diaion™SP70, Diaion™ SP 205, Diaion™ SP 206, Diaion™ SP 207, Diaion™ SP 700,Diaion™ SP 800, Diaion™ SP 825, Diaion™ SP 850, Diaion™ SP 875, Diaion™HP 1MG, Diaion™ HP 2MG, Diaion™ CHP 55A, Diaion™ CHP 55Y, Diaion™ CHP20A, Diaion™ CHP 20Y, Diaion™ CHP 2MGY, Diaion™ CHP 20P, Diaion™ HP20SS, Diaion™ SP 20SS, Diaion™ SP 207SS. Purolite Company: Purosorb™ AP250 and Purosorb™ AP 400, and Kaneka Corp. Lixelle beads.

Various proteins may be adsorbed by the composition of the instantdisclosure. Some of these proteins and their molecular weights are shownin the table below.

Protein Molecular Weight (Da) PAF (Platelet Activating Factor) 524bilirubin 548.6 heme b 616.5 MIP-1alpha 8,000 Complement C5a 8,200Complement C3a 9,089 IL-8 9,000 S100B (dimerizes) 10,000 β-2microglobulin 11,800 Procalcitonin 13,000 Phospholipase A2, secretoryPLA2 type I 14,000 pancreatic PLA2G2A 16,083 IL-7 17,400 Myoglobin17,699 Trypsin-human pancreas 23,300 IL-6 23,718 Toxic shock syndrometoxin 1 (TSST-1 24,000 Enterotoxin B, S aureus 24,500 HMGB1 24,894Interferon gamma 25,000 Chymotrypsin 25,000 Elastase (neutrophil) 25,000Trypsin 26,488 PF4 27,100 Enterotoxin A, S. aureus 27,800 alpha toxinA&B, S. aureus 28,000 PCNA, proliferating cell nuclear antigen 29,000Arginse I 35,000 Carboxypeptidase A 35,000 Thrombin 36,700 alpha-1antitrypsin 44,324 TNF-alpha 52,000 Activated Protein C 56,200 Amylase57,000 hemopexin 57,000 alpha-1 antichymotrypsin 55,000-68,000 Diptheriatoxoid 62,000 hemoglobin, oxy 64,000 Pseudomonas Exotoxin A 66,000ShigaToxin (A 32 kDa, 5 × B 7.7 kDa) 69,000 Calpain-1 (humanerythrocytes) 112,00 C reactive Protein (5 × 25 kDa) 115,000Myeloperoxidase (neutrophils) 150,000 Immunoglobulin G IgG 150,000 NOSsynthase 150,000 Immunoglobulin A IgA 162,000 Immunoglobulin E (IgE)190,000 Immunoglobulin M IgM 950,000

The following examples are intended to be exemplary and non-limiting.

Example 1: Base Sorbent Synthesis CY12004, CY15042, CY15044, CY15045,and CY15077

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Reactor Setup; a 4-neck glass lid was affixed to a 3000 mL jacketedcylindrical glass reaction vessel using a stainless steel flange clampand PFTE gasket. The lid was fitted with a PFTE stirrer bearing, RTDprobe adapter, and water-cooled reflux condenser. A stainless steelstirring shaft having five 45° agitators was fit through the stirrerbearing and inserted into a digital overhead stirrer. An RTD probe wasfit through the corresponding adapter, and connected to a PolyStatcirculating heating and chilling unit. Compatible tubing was used toconnect the inlet and outlet of the reaction vessel jacket to theappropriate ports on the Poly Stat. The unused port in the lid was usedfor charging the reactor and was plugged at all other times.

Polymerization; Aqueous phase and organic phase compositions are shownbelow, in Table I and Table II, respectively. Ultrapure water was splitinto approximately equal parts in two separate Erlenmeyer flasks eachcontaining a PFTE coated magnetic stir bar. Poly(vinyl alcohol) (PVA),having a degree of hydrolysis of 85.0 to 89.0 mol percent and aviscosity of 23.0 to 27.0 cP in a 4% aqueous solution at 20° C., wasdispersed into the water in the first flask and heated to 80° C. on ahot plate with agitation. Salts (see Table 1, MSP, DSP, TSP and Sodiumnitrite) were dispersed into the water in the second flask and heated to80° C. on a hot plate with agitation. Circulation of heat transfer fluidfrom the PolyStat through the reaction vessel jacket was started, andfluid temperature heated to 60° C. Once PVA and salts dissolved, bothsolutions were charged to the reactor, one at a time, using a glassfunnel. The digital overhead stirrer was powered on and the rpm set to avalue to form appropriate droplet sizes upon organic phase addition.Temperature of the aqueous phase in the kettle was set to 70° C. Theorganic phase was prepared by adding benzoyl peroxide (BPO) to thedivinylbenzene (DVB) and styrene in a 2 L Erlenmeyer flask and swirlinguntil completely dissolved. 2,2,4-trimethylpentane and toluene wereadded to the flask, which was swirled to mix well. Once the temperatureof the aqueous phase in the reactor reached 70° C., the organic phasewas charged into the reactor using a narrow-necked glass funnel.Temperature of the reaction volume dropped upon the organic addition. Atemperature program for the PolyStat was started, heating the reactionvolume from 60 to 77° C. over 30 minutes, 77 to 80° C. over 30 minutes,holding the temperature at 80° C. for 960 minutes, and cooling to 20° C.over 60 minutes. 1-Vinyl-2-pyrrolidinone (VP) was added dropwise viaglass separatory funnel once the reaction reached identity point,approximately one hour after the reaction temperature reached 80° C.Note: the temperature program for preparation of polymer CY15042 wasdifferent, proceeding as follows; reaction volume heated from 55 to 62°C. over 30 minutes, 62 to 65° C. over 30 minutes, held at 65° C. for1320 minutes, heated from 65 to 82° C. over 30 minutes, 82 to 85° C.over 30 minutes, held at 85° C. for 60 minutes, then cooled to 20° C.over 60 minutes.

TABLE I Aqueous Phase Composition Reagent Mass (g) Ultrapure water1500.000 Poly(vinyl alcohol) (PVA) 4.448 Monosodium phosphate (MSP)4.602 Disodium phosphate (DSP) 15.339 Trisodium phosphate (TSP) 9.510Sodium nitrite 0.046 Total 1533.899

TABLE II Organic Phase Compositions CY12004 CY15042 CY15044 CY15045CY15077 Reagent Mass (g) Mass (g) Mass (g) Mass (g) Mass (g)Divinylbenzene, 63% (DVB) 508.751 451.591 386.284 386.284 498.383Styrene 0.000 0.00 374.118 374.118 0.000 2,2,4-trimethylpentane(Isooctane) 384.815 125.800 271.210 271.210 482.745 Toluene 335.004712.869 235.725 235.725 222.404 Benzoyl peroxide, 98% (BPO) 3.816 18.4325.703 5.703 3.738 1-Vinyl-2-pyrrolidinone (VP) 151.578 0.000 0.000167.288 0.000 Total (excluding BPO and VP) 1228.571 1290.260 1267.3371267.337 1203.532

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Work-up; reaction volume level in the reactor was marked. Overheadstirrer agitation was stopped, residual liquid siphoned out of thereactor, and the reactor filled to the mark with ultrapure water at roomtemperature. Overhead stirrer agitation was restarted and the slurryheated to 70° C. as quickly as possible. After 30 minutes, agitation wasstopped and residual liquid siphoned out. Polymer beads were washed fivetimes in this manner. During the final wash, the slurry temperature wascooled to room temperature. After the final water wash, polymer beadswere washed with 99% isopropyl alcohol (IPA) in the same manner. 99% IPAwas siphoned out and replaced with 70% IPA before transferring theslurry into a clean 4 L glass container. Unless noted otherwise, on anas-needed basis the polymer was steam stripped in a stainless steel tubefor 8 hours, rewet in 70% IPA, transferred into DI water, sieved toobtain only the portion of beads having diameters between 300 and 600μm, and dried at 100° C. until no further weight loss on drying wasobserved.

Cumulative pore volume data, measured by nitrogen desorption isotherm,for polymers CY12004, CY15042, CY15044, and CY15045, are presentedbelow, in Tables III, IV, V, and VI, respectively. Cumulative porevolume data, measured by mercury intrusion porosimetry, for polymerCY15077 is presented in Table VII, below.

TABLE III Nitrogen Desorption Data for CY12004 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 1221.6-868.1 985.2149834 0.009113091 868.1-751.9 801.4105771 0.019081821 751.9-661.5700.749642 0.032021618 661.5-613.5 635.6650389 0.048206769 613.5-568.5589.2088599 0.067981224 568.5-509.8 535.8385194 0.114704165 509.8-456.1479.8625277 0.214714265 456.1-418.7 435.7117054 0.311269356 418.7-374.6394.0534583 0.455991378 374.6-330.2 349.456374 0.579735461 330.2-319.6324.7147611 0.612988132 319.6-281.8 298.1620033 0.708072633 281.8-273.9277.7142728 0.73291244 273.9-256.6 264.6494358 0.777049805 256.6-237.0245.9517985 0.830089884 237.0-225.7 231.0229263 0.857298007 225.7-215.6220.375968 0.88145223 215.6-145.5 166.3375231 1.066971104 145.5-104.6117.8539174 1.181204175 104.6-84.4  92.0541661 1.241569291 84.4-71.476.67121175 1.285618005 71.4-60.9 65.20679768 1.326059561 60.9-52.756.07123392 1.360787093 52.7-46.5 49.12518253 1.389258246 46.5-41.343.53851295 1.416541075 41.3-37.1 38.91936166 1.445235862

TABLE IV Nitrogen Desorption Data for CY15042 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 2011.0-633.1 751.380276 0.003621266 633.1-424.8 488.0919378 0.006317461 424.8-418.5421.593936 0.006912678 418.5-353.5 380.29179 0.008267096 353.5-280.4308.2300243 0.011094129 280.4-275.4 277.8814342 0.01168737 275.4-249.5261.1230419 0.012721633 249.5-209.1 225.543664 0.015611261 209.1-206.8207.9070897 0.016388077 206.8-137.5 157.790999 0.442556595 137.5-98.1 110.7933773 0.765560391 98.1-81.8 88.28728758 0.845836735 81.8-67.372.96250925 0.911182647 67.3-57.7 61.63744463 0.954008444 57.7-50.353.43111186 0.983515641 50.3-44.4 46.93705679 1.010486042 44.4-38.641.02620024 1.037817277 38.6-34.5 36.30857144 1.058861412 34.5-30.932.48566551 1.08400665 30.9-27.3 28.85395017 1.10131894 27.3-24.325.59611525 1.12576046 24.3-22.3 23.18199338 1.143118464 22.3-19.620.72009386 1.167009752 19.6-17.4 18.32182238 1.190109864

TABLE V Nitrogen Desorption Data for CY15044 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 2529.6-789.0 936.1742201 1.75877E−06 789.0-446.0 526.203721 0.000135623 446.0-219.6260.7379647 0.002068559 219.6-213.4 216.3756282 0.004663144 213.4-205.7209.3598959 0.0088853 205.7-144.7 164.0510277 0.131650053 144.7-99.6 113.2793455 0.294709491 99.6-82.1 88.98089675 0.331539838 82.1-71.475.89033961 0.34527909 71.4-60.0 64.52630192 0.360216738 60.0-52.855.83732662 0.367929549 52.8-46.8 49.32751384 0.373710394 46.8-41.443.66300585 0.378313283 41.4-37.2 39.02724789 0.38481289 37.2-33.234.8920748 0.391803441 33.2-30.0 31.34913535 0.393761301 30.0-27.328.49102813 0.394422444 27.3-24.7 25.83440471 0.396180539 24.7-22.323.34690716 0.401510134 22.3-19.8 20.83368622 0.40782788 19.8-17.518.45917969 0.416568116

TABLE VI Nitrogen Desorption Data for CY15045 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 1277.7-542.6 649.560333 0.000489722 542.6-213.2 252.9981774 0.000667721 213.2-206.9209.9696024 0.001419558 206.9-141.9 161.6476715 0.261729457 141.9-106.3118.498425 0.346563251 106.3-84.0  92.17838423 0.37856771 84.0-71.876.76600632 0.393497452 71.8-62.4 66.31374327 0.404409264 62.4-53.657.17863111 0.411077722 53.6-48.0 50.38372676 0.416000386 48.0-42.544.81172299 0.421626585 42.5-38.3 40.08447096 0.428067208 38.3-34.436.07215077 0.431303175 34.4-31.5 32.76081107 0.433543649 31.5-26.327.29321095 0.440720595 26.3-23.8 24.89263623 0.44207166 23.8-21.322.31849785 0.443967237 21.3-19.1 19.99937462 0.45436982 19.1-16.117.16801839 0.47745598

TABLE VII Mercury Intrusion Data for CY15077 Pore size Diameter (A)Cumulative Intrusion (mL/g) 226299.0625 3.40136E−30 213166.07810.001678752 201295.1563 0.002518128 172635.8125 0.004364755 139538.06250.007554384 113120.7813 0.011919139 90542.36719 0.01645177 78733.257810.0203129 72446.375 0.022327403 60340.40234 0.027867284 48343.839840.035327822 39009.13672 0.040918175 32136.4082 0.04899035 25330.656250.063195683 20981.51563 0.079529688 16219.86426 0.108860672 13252.412110.141730919 10501.53613 0.193969816 8359.911133 0.262399256 6786.301270.345866203 5538.122559 0.438174427 4337.931152 0.563276172 3501.6748050.681870878 2838.742188 0.804727197 2593.016846 0.865813017 2266.6889650.938610673 1831.041748 1.056586146 1509.850708 1.163395643 1394.0061041.21002543 1294.780151 1.257248282 1207.692627 1.293158531 1131.8609621.326992273 1065.099976 1.35812819 953.1816406 1.405935764 884.03588871.445426106 823.5491333 1.478719592 770.9108276 1.510579824 722.47247311.537048101 684.6119995 1.564400196 672.187561 1.581117511 636.78857421.60271585 604.7248535 1.621845484 558.1287231 1.651492 518.26245121.678913713 483.5536499 1.708594561 453.5110779 1.735918999 426.99984741.755934 403.1251526 1.783603072 382.7776794 1.793849826 362.71624761.817784309 342.3734436 1.838774562 330.1105042 1.851493955 315.52380371.869742155 302.2973938 1.885128617 290.2946777 1.895119786 279.12466431.912378907 268.7442627 1.924305081 259.1106873 1.936048627 241.87377931.955100656 226.7678223 1.972970247 213.3626251 1.988123298 201.49081422.007521152 194.9888611 2.022114754 188.9506989 2.033871174 180.5829012.035052776 172.8530121 2.050720692 164.9621735 2.062945843 157.81106572.071056128 151.1540375 2.082133055 143.9185333 2.096480608 138.46705632.106938839 132.8492737 2.119287968 129.5760345 2.126605988 126.54386142.126605988 124.2635574 2.132267475 120.8976135 2.141504765 117.37922672.150759459 114.791893 2.154810667 111.9475937 2.162935257 108.88300322.167646885 106.6480179 2.174062729 104.5217743 2.179908991 102.42951972.179908991 100.1580353 2.182951927 98.29322052 2.184018135 96.448226932.191127539 94.42159271 2.198545218 91.52587891 2.209161043 89.258079532.209312439 87.0777359 2.215425491 85.42358398 2.221472025 83.626129152.232139587 82.11174011 2.237514496 79.91614532 2.239231586 78.014625552.239560127 76.19993591 2.239560127 75.09249115 2.239560127 73.412010192.239560127 72.23709869 2.240245819 71.09960175 2.242422104 69.863014222.243849993 68.40761566 2.257676363 67.13697815 2.259181261 66.033592222.266284466 65.08189392 2.270181179 64.04368591 2.272682428 62.384902952.280714512 61.32764053 2.280714512 60.30379868 2.287917852 59.413703922.287917852 58.54679489 2.293802738 57.79866409 2.297607183 56.889778142.299046278 55.9213295 2.302111387 54.98665237 2.303381443

Example 2: Polymer Modification CY15087

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

N-vinylpyrrolidone functionalization; base polymer, CY15077, was notsteam stripped or sieved prior to functionalization. Two 99% IPA washesat 50° C. were completed during workup for the base polymer, as opposedto one wash at RT. Following IPA washes, the polymer was washed threetimes with an excess of DI water. Wetted CY15077 polymer beads wereadded to a 1 L jacketed glass reaction kettle, fitted with a Tefloncoated agitator, containing 450 mL DI water, 50.0 g N-vinylpyrrolidonemonomer, and 1.5 g sodium persulfate. The reaction was allowed toproceed for 24 hours at 75° C., with agitation speed set to 100 RPM.Upon completion the polymer beads were washed five times with 500 mL DIwater at 70° C., steam stripped in a stainless steel tube for 8 hours,rewet in 70% IPA, transferred into DI water, sieved to obtain only theportion of beads having diameters between 300 and 600 μm, and dried at100° C. until no further weight loss on drying was observed. The yieldwas 95.5 g of polymer CY15087. Atomic concentrations measured by XPS,and cumulative pore volume data measured by mercury intrusionporosimetry, are shown in Tables VIII and IX, respectively.

TABLE VIII Atomic Concentrations (in %) for CY15077 and CY15087 PolymerCondition C N O CY15077 Bead 96.2 0.0 3.8 CY15077 Ground 98.6 0.0 1.4CY15087 Bead 95.5 0.4 4.2 CY15087 Ground 98.3 0.2 1.5

TABLE IX Mercury Intrusion Data for CY15087 Pore size Diameter (A)Cumulative Intrusion (mL/g) 226275.6875 3.003E−30 213126.625 0.001333927201250.5938 0.002964283 172601.8438 0.005928566 139532.5469 0.009189277113124.3359 0.012449989 90545.25 0.015710698 78739.35156 0.01748926972432.5625 0.01897141 60333.77734 0.021935694 46762.60547 0.02679563939173.96094 0.03074207 31808.34375 0.034442116 25357.64648 0.04002706720929.94141 0.046409778 16182.15234 0.056623131 13255.21973 0.06579688910561.28809 0.080750667 8353.926758 0.105692402 6778.929199 0.1386706835543.002441 0.177410021 4342.263672 0.24024339 3502.678711 0.3080583212839.226807 0.388105094 2591.51416 0.428066701 2267.699951 0.481548221831.208252 0.570007741 1510.12561 0.655585647 1394.226563 0.6961807011294.746582 0.729135811 1208.07251 0.76245892 1132.023804 0.7959909441065.684937 0.815372229 953.989502 0.855566621 883.8703613 0.871785223823.4996338 0.921781898 771.3513794 0.949763238 722.1901245 1.018806458684.8914185 1.027466536 671.8579712 1.033001781 636.456604 1.044957519604.6593018 1.05753231 557.9059448 1.079107881 518.4785156 1.102458835483.8456726 1.127018452 453.9489746 1.151340365 426.8711243 1.174746156402.8918152 1.194709539 382.4490967 1.213674426 360.680481 1.231868267342.5672302 1.252067924 329.8339539 1.267953753 315.4637756 1.28668797302.4020996 1.299176812 290.331665 1.314114213 279.2361145 1.322446585268.7993164 1.34148562 259.2027283 1.349915743 241.8540192 1.363333344226.7354431 1.38415575 213.408844 1.386666298 201.5056763 1.411639214194.9947357 1.426415801 188.935318 1.428328514 180.6179199 1.441128492172.8575745 1.453100324 164.9869385 1.464205742 157.740097 1.473819733151.1829987 1.486423731 143.9502716 1.499343991 138.4791107 1.509965897132.8890839 1.522242427 129.5950317 1.529255748 126.493248 1.529255748124.2660522 1.53686142 120.8921432 1.543375134 117.3944702 1.549948096114.7864304 1.558065772 111.9504318 1.56092155 108.9145203 1.564850807106.6669846 1.571887255 104.5330276 1.574593782 102.4421844 1.584572434100.1668015 1.591516852 98.28172302 1.594149351 96.44982147 1.59504282594.43471527 1.595328212 91.55084229 1.595610261 89.27562714 1.60425078987.08631134 1.61047101 85.43348694 1.616541862 83.63105011 1.62080562182.10086823 1.627643347 79.91345978 1.629765868 78.01348877 1.63120782476.20350647 1.63190341 75.09172821 1.634262919 73.41147614 1.63839113772.23751831 1.642881751 71.10028076 1.646320224 69.861763 1.64873695468.40744019 1.655003667 67.13788605 1.662294388 66.03204346 1.66740560565.08184814 1.670548201 64.04498291 1.671463728 62.38602829 1.67300248161.32709885 1.673002481 60.30479813 1.673002481 59.41309738 1.67300248158.54596329 1.673002481 57.799366 1.673613429 56.88968277 1.67361342955.92052078 1.677541733 54.98633194 1.677541733

Example 3: Polymer Modification CY15100 and CY15102

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Sulfonation procedure; dried base polymer was added to a 1 L jacketedglass reactor, which was equipped with a Teflon coated agitator. Amixture of concentrated sulfuric acid (98%) and fuming sulfuric acid(20% SO₃ in sulfuric acid) was added to the reactor containing basepolymer. The reaction was carried out at 90° C. for 24 hours, withconstant agitation at 100 RPM.

Work-up; the reaction volume was allowed to cool to room temperature(RT), and was slowly added into a chemical glass beaker with an excessof at least 1 L ice cold DI water. Sulfonated polymer was washed withexcess DI water at RT until the supernatant reached a neutral pH. Theresulting polymer was then treated with 100 mL 1M NaOH_((aq)) for 1 hourat RT to convert polymer bound ˜SO₃H into ˜SO₃Na groups. Polymer waswashed again with an excess of DI water at RT until the supernatantreached a neutral pH, then dried in an oven at 100° C. until no furtherloss on drying was observed. The dried ˜SO₃Na functional polymer yieldwas measured. Reaction compositions for CY15100 and CY15102 are providedin Table X. Table XI displays atomic concentrations for polymersCY15100, CY15102, and CY15087, as measured by XPS. Log differential porevolume plots are presented in FIGS. 1, 2, and 3, and cumulative porevolume data are presented in Tables XII, XIII, and XIV. Wheninterpreting pore structure data obtained from nitrogen desorptionisotherm or mercury intrusion porosimetry using dried polymer as thesample, it is important to consider that pore size may change uponswelling of sulfonated poly(styrene-co-divinylbenzene) porous beads oncewetted in solution. In addition to potential changes in pore structure,the bead size may also change upon transition from dry to swollen state.This phenomenon was evaluated in “Preparation and Evaluation ofDifferently Sulfonated Styrene-Divinylbenzene Cross-Linked CopolymerCationic Exchange Resins as Novel Carriers for Drug Delivery”, publishedin AAPS PharmSciTech June 2009; 10(2): 641-648.

Thrombogenicity was measured by the uPTT assay in which materials werecompared to the negative control (plasma alone), positive control (glassbeads) and reference beads to determine the degree of contact activationactivity. In the uPTT assay, the % change in clot formation over time ascompared to the reference materials was determined, then groupedaccording to: <25% activators, 25-49% moderate activators, 50-74% mildactivators, 75-100% minimal and >100% non-activators of the intrinsiccoagulation pathway. Polymer CY15100, 82%, was a minimal activator.

TABLE X Modification Compositions for CY15100 and CY15102 CY15100CY15102 Base Polymer CY15045 CY15087 Mass Base Polymer (g) 220.0 80.0Mass Concentrated Sulfuric Acid (g) 950.0 550.0 Mass Fuming SulfuricAcid (g) 50.0 30.0 Yield Dry Modified Polymer (g) 355.5 204.6

TABLE XI Atomic Concentrations (in %) for CY15100, CY15102, and CY15087Polymer Condition C N O Na S CY15100 Bead 65.7 0.2 20.5 8.0 5.7 CY15102Bead 71.6 0.5 17.2 6.6 4.1 CY15087 Bead 95.5 0.4 4.2 0.0 0.0 CY15087Ground 98.3 0.2 1.5 0.0 0.0

TABLE XII Nitrogen Desorption Data for CY15100 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 2072.6-552.5 648.562383 0.000416099 552.5-354.3 410.9223564 0.000905416 354.3-337.5345.4980521 0.001122701 337.5-311.7 323.5292132 0.001561729 311.7-288.8299.3515093 0.001919004 288.8-272.0 279.8911375 0.002345465 272.0-252.4261.4265539 0.002783018 252.4-239.0 245.303922 0.003244227 239.0-225.7231.9701343 0.004052829 225.7-212.7 218.7962082 0.005280802 212.7-204.5208.4059706 0.007418375 204.5-131.6 152.2941936 0.087124099 131.6-99.3 110.8101833 0.170472908 99.3-72.3 81.43320551 0.20555251 72.3-62.266.46726774 0.212857437 62.2-52.1 56.21812708 0.218554756 52.1-45.748.42881742 0.221509707 45.7-39.4 42.03869424 0.223879096 39.4-34.536.58507436 0.225521077 34.5-29.1 31.32500867 0.230015257 29.1-25.126.83485239 0.230195589 25.1-22.0 23.36857621 0.230286279 22.0-19.420.51213112 0.232863812

TABLE XIII Nitrogen Desorption Data for CY15102 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 1598.8-1238.81372.941344 0.026272341 1238.8-946.4  1053.220361 0.092081573946.4-758.1 831.0482586 0.194131921 758.1-677.1 712.8857859 0.258283006677.1-529.4 584.7345957 0.38744334 529.4-485.2 505.2928332 0.431560876485.2-443.2 462.211297 0.481712669 443.2-396.6 417.205311 0.529431372396.6-361.7 377.5005059 0.571368548 361.7-324.6 341.0804153 0.616836751324.6-296.1 308.9881056 0.653318693 296.1-271.6 282.7268533 0.684779469271.6-256.8 263.7792955 0.704908544 256.8-239.4 247.4475287 0.727833901239.4-230.2 234.5702219 0.739926683 230.2-217.1 223.2037828 0.756372211217.1-206.8 211.680438 0.769442061 206.8-140.7 160.5985991 0.863794835140.7-106.0 118.0466801 0.922556622 106.0-82.8  91.24150017 0.96803966182.8-68.2 73.90381313 1.001829887 68.2-60.9 64.05281388 1.02092002360.9-52.5 55.98109194 1.044868856 52.5-46.3 48.94597942 1.06524739746.3-41.2 43.35983259 1.084233305 41.2-37.0 38.81504369 1.10645690837.0-33.1 34.78421912 1.129603729 33.1-30.0 31.33519542 1.14621880130.0-27.3 28.4688972 1.162517069 27.3-24.6 25.75215358 1.18204862824.6-22.4 23.33791231 1.201310022 22.4-19.8 20.89212489 1.22914870619.8-17.6 18.54892595 1.260822457

TABLE XIV Mercury Intrusion Data for CY15102 Pore size Diameter (A)Cumulative Intrusion (mL/g) 226247.25 3.02E−30 213156.0625 0.000893837201297.1875 0.002383566 172619.2656 0.004320214 139526.7344 0.006107889113150.6484 0.007448644 90544.85156 0.009236319 78737.24219 0.01013015672447.07031 0.011172966 60339.52344 0.012066803 49074.61719 0.01206680338783.65625 0.012066803 32031.35742 0.012456137 25154.1582 0.0155003720919.94336 0.01550037 16226.36035 0.016433783 13231.0293 0.01806502610569.24219 0.020413134 8346.358398 0.023545867 6777.795898 0.0275560935545.635742 0.032167129 4347.45166 0.039555997 3496.898926 0.0492774362839.973145 0.057190847 2592.47998 0.06178461 2267.395264 0.0716473091831.758789 0.089788206 1510.39563 0.112907536 1394.068237 0.1257442531294.699707 0.136810422 1207.551147 0.147966579 1132.260498 0.1595866081065.672974 0.171025708 954.0095215 0.191800222 884.2581177 0.20811981823.8370972 0.228217274 771.1380615 0.239915013 721.8734131 0.275565475684.4716797 0.281177133 672.791748 0.283745468 636.3512573 0.295114249605.4035034 0.309263676 558.758606 0.326112717 518.5050049 0.352752388483.7310181 0.367008656 453.6919861 0.390547335 426.9628296 0.407471895403.0959778 0.4232741 382.8546753 0.444355428 362.905426 0.463873088342.0473328 0.487040371 329.7276001 0.504495382 315.7310791 0.522837102302.3917236 0.545027971 290.2372131 0.567096949 279.1113586 0.588691056268.6489563 0.608853817 259.2150879 0.635331511 241.9123993 0.710671127226.7029877 0.774290979 213.3559113 0.867704988 201.5307922 0.867704988195.0246887 0.867704988 188.9438019 0.867704988 180.6033783 0.867704988172.8410034 0.869671643 164.969101 0.869671643 157.8126526 0.87475878151.1803131 0.905465066 143.936264 0.909094393 138.4554596 0.931292474132.8584442 0.938616037 129.575531 0.938616037 126.4766693 0.971493781124.2657852 0.971493781 120.9015427 0.972762465 117.374855 0.977469385114.7828751 0.981295645 111.9444351 0.981295645 108.8816452 0.981295645106.6592331 0.986702561 104.5428238 0.996097863 102.4358368 1.000003457100.1722946 1.003374338 98.26839447 1.006461024 96.44637299 1.00896668494.41146851 1.012030125 91.54938507 1.015347958 89.25726318 1.01844024787.0788269 1.021567345 85.42123413 1.024644852 83.62944031 1.02823948982.1011734 1.02980864 79.91355133 1.032312155 78.00926208 1.03494870776.20082092 1.037501097 75.09120178 1.039880157 73.4092865 1.04204273272.23842621 1.043176413 71.09993744 1.047091961 69.86208344 1.04725861568.40840912 1.049208641 67.1362381 1.05278945 66.0329895 1.0527894565.08166504 1.053350925 64.04417419 1.054639339 62.38519287 1.05590236261.32834625 1.060090899 60.30381012 1.062460899 59.41312408 1.06342089258.54793549 1.064275384 57.79902267 1.066532493 56.88972473 1.06811249355.92105865 1.072528958 54.9865036 1.072528958

Example 4: Polymer Modification CY14144 and CY15101

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Sulfonation procedure; dried base polymer was mixed with glacial aceticacid in a 500 mL glass reactor equipped with a Teflon coated mechanicalagitator, and heated to 50° C. with agitation set to 100 RPM. A mildsulfonating agent was prepared by adding acetic anhydride (99%) to a 100mL chemical glass beaker, cooled in an ice bath, and slowly addingconcentrated sulfuric acid (98%) over 30 minutes. Temperature of themixture was monitored and maintained between 15-40° C. by replenishingthe ice bath. After completion of the sulfuric acid addition, thereddish-brown viscous liquid was kept at RT for 1 hour, and then slowlyadded to the reactor. The reaction was allowed to proceed for aspecified amount of time.

Work-up; the reaction volume was allowed to cool to room temperature(RT), and was slowly added into a chemical glass beaker with an excessof at least 1 L ice cold DI water. Sulfonated polymer was washed withexcess DI water at RT until the supernatant reached a neutral pH. Theresulting polymer was then treated with 100 mL 1M NaOH_((aq)) for 1 hourat RT to convert polymer bound ˜SO₃H into ˜SO₃Na groups. Polymer waswashed again with an excess of DI water at RT until the supernatantreached a neutral pH, then dried in an oven at 100° C. until no furtherloss on drying was observed. The dried ˜SO₃Na functional polymer yieldwas measured. Reaction compositions for polymers CY14144 and CY15101 areshown in Table XV, below. Atomic concentrations determined by XPS forpolymers CY14144, CY12004, CY15101, and CY15087 are presented below, inTable XVI. FIGS. 4, 5, and 6 show plots of log differential pore volumefor each of the modified polymers described above. Cumulative porevolume data are shown below in Tables XVII, XVIII, and XIX.

Thrombogenicity was measured by the uPTT assay in which materials werecompared to the negative control (plasma alone), positive control (glassbeads) and reference beads to determine the degree of contact activationactivity. In the uPTT assay, the % change in clot formation over time ascompared to the reference materials was determined, then groupedaccording to: <25% activators, 25-49% moderate activators, 50-74% mildactivators, 75-100% minimal and >100% non-activators of the intrinsiccoagulation pathway. Polymer CY15101, 88%, was a minimal activator.

TABLE XV Modification Compositions for CY14144 and CY15101 CY14144CY15101 Base Polymer CY12004 CY15087 Mass Base Polymer (g) 11.7 80.5Volume Glacial Acetic Acid (mL) 75 400 Mass Acetic Anhydride (g) 15.5125.0 Mass Concentrated Sulfuric Acid (g) 10.0 80.0 Reaction Time (hr) 12 Yield Dry Modified Polymer (g) 15.2 103.4

TABLE XVI Atomic Concentrations (in %) for CY14144, CY12004, CY15101 andCY15087 Polymer Condition C N O Na S CY14144 Ground 87.0 0.0 8.7 2.5 1.8CY12004 Bead 88.7 3.4 7.9 0.0 0.0 CY12004 Ground 95.0 0.4 4.7 0.0 0.0CY15101 Bead 93.3 0.7 5.5 0.4 0.1 CY15087 Bead 95.5 0.4 4.2 0.0 0.0CY15087 Ground 98.3 0.2 1.5 0.0 0.0

TABLE XVII Nitrogen Desorption Data for CY14144 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 1316.7-872.8 1006.357045 0.00765904 872.8-760.0 808.4322272 0.01624896 760.0-683.5717.5974884 0.028034508 683.5-625.7 651.9899544 0.04390322 625.7-580.4601.3250683 0.063539075 580.4-506.1 537.9818896 0.171853178 506.1-449.5474.3191955 0.315569993 449.5-395.6 418.9973346 0.494055963 395.6-367.4380.4061561 0.562885137 367.4-336.7 350.6379611 0.677258819 336.7-297.9314.7821036 0.775586161 297.9-293.1 295.4222209 0.799779319 293.1-271.2281.2260787 0.844974101 271.2-254.9 262.5094171 0.885670627 254.9-241.5247.8251105 0.914384164 241.5-229.9 235.3841146 0.939107638 229.9-218.4223.8303393 0.963836661 218.4-143.9 165.3652577 1.129471233 143.9-105.5118.3403917 1.219663568 105.5-85.5  93.18473659 1.269908393 85.5-70.276.1345754 1.313809596 70.2-59.9 64.12468772 1.346563035 59.9-51.855.16793458 1.375536168 51.8-45.4 48.0976978 1.401122481 45.4-40.242.43389408 1.424635177 40.2-36.1 37.88651456 1.449718809 36.1-32.133.79019728 1.477483715 32.1-28.9 30.28159181 1.497726602 28.9-26.327.45536434 1.516706872 26.3-23.6 24.73777781 1.540207545 23.6-21.122.17739744 1.565080566 21.1-19.0 19.93703511 1.59109954 19.0-16.517.51596978 1.630846881

TABLE XVIII Nitrogen Desorption Data for CY15101 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 1633.5-1308.61434.817706 0.015869195 1308.6-938.9  1063.085479 0.076011287938.9-822.1 872.5400938 0.113427572 822.1-664.4 726.153686 0.190492729664.4-541.2 589.8410779 0.271968628 541.2-495.4 516.2061221 0.306797766495.4-452.4 471.8824199 0.342922234 452.4-411.9 430.1606805 0.374728484411.9-373.7 390.8559715 0.406903208 373.7-338.1 354.0440222 0.436879429338.1-306.8 320.8627294 0.464287882 306.8-282.0 293.2737773 0.487609004282.0-258.7 269.2384186 0.507679709 258.7-244.7 251.276503 0.519925622244.7-229.0 236.3042502 0.53405423 229.0-216.9 222.6261956 0.545016489216.9-207.3 211.8651212 0.55492914 207.3-144.0 163.6239028 0.618758477144.0-103.4 116.4570854 0.668697015 103.4-85.6  92.59846579 0.69426377885.6-72.1 77.55016135 0.716487235 72.1-60.7 65.25664927 0.7373396760.7-53.0 56.18831893 0.752958019 53.0-46.6 49.26725758 0.76734478446.6-41.5 43.64795708 0.78015016 41.5-37.3 39.09541737 0.79498198937.3-33.3 35.01993833 0.810085989 33.3-30.2 31.57107231 0.820134730.2-27.5 28.71043051 0.829560837 27.5-24.8 25.99331273 0.83997554424.8-22.6 23.59379328 0.8493402 22.6-20.0 21.1163566 0.86190814320.0-17.8 18.73336683 0.874402144

TABLE XIX Mercury Intrusion Data for CY15101 Pore size Diameter (A)Cumulative Intrusion (mL/g) 226247.25 3.367E−30 213156.0625 0.001661795201297.1875 0.002658872 172619.2656 0.005317744 139526.7344 0.007976616113150.6484 0.009638411 90544.85156 0.012297283 78737.24219 0.01412525772447.07031 0.015620873 60339.52344 0.017947385 49556.56641 0.0194987838738.37109 0.021929506 31002.00586 0.023903539 25333.7832 0.0261679220724.62109 0.029177248 16168.99121 0.033409968 13230.375 0.03776564110563.43555 0.044586275 8346.731445 0.054462213 6776.340332 0.06665465536.147949 0.083998173 4342.036621 0.107802272 3501.501953 0.1374850422837.420654 0.177576199 2594.672363 0.200087309 2269.617432 0.2334893791831.204224 0.295208424 1510.503906 0.360582143 1395.643555 0.3929020171293.973755 0.421268374 1207.494141 0.447410613 1131.894531 0.472416581065.193237 0.49649471 953.9039307 0.535679519 884.3017578 0.568524599823.786377 0.597846568 771.5706177 0.616960466 722.1925049 0.691450536684.2458496 0.697761118 672.2320557 0.703246117 636.7992554 0.713504672604.4926758 0.726847529 558.8725586 0.746505737 517.9966431 0.774387836483.9524536 0.799027622 453.7037354 0.824069798 426.9303894 0.846621335403.1401672 0.898474514 382.6773987 0.91877532 362.9386292 0.946397841342.2199707 0.946397841 330.153656 0.953894079 315.6123962 0.954481184302.6812439 0.954481184 290.4436646 0.967425823 279.009491 0.974567354268.8323975 0.974567354 259.2565308 0.974567354 241.9353333 1.028741002226.8330078 1.048289418 213.444046 1.065926313 201.5080414 1.074228048195.0001221 1.095143437 188.9437103 1.106776357 180.6530914 1.11556828172.9412994 1.127364159 164.9789429 1.139592171 157.7405396 1.150992036151.1612091 1.161784291 143.9489746 1.172875285 138.4779053 1.185242534132.8603821 1.194525123 129.5736542 1.200321555 126.4793472 1.207967401124.2483292 1.213427901 120.9080048 1.221876502 117.3827286 1.223723292114.771225 1.23161757 111.937149 1.237899184 108.9081039 1.239180923106.6535568 1.245096564 104.5474396 1.24916625 102.455368 1.25267899100.1680145 1.261325955 98.2784729 1.261325955 96.45231628 1.26788520894.40316772 1.274962544 91.53180695 1.279593945 89.26702118 1.28591597187.08314514 1.285915971 85.42582703 1.287682652 83.6335144 1.2940089782.10058594 1.300267935 79.91345978 1.30387032 78.01080322 1.30826437576.19985962 1.313777924 75.09228516 1.318249345 73.41210175 1.32150864672.23653412 1.323805094 71.09803772 1.32500124 69.86273193 1.33416771968.40810394 1.336985707 67.13769531 1.340026617 66.03487396 1.34002661765.0819931 1.340224981 64.04338074 1.340224981 62.38589478 1.34669029761.32817841 1.358168244 60.30670166 1.358168244 59.41316605 1.35853290658.54763031 1.358532906 57.79816818 1.358532906 56.88824844 1.35853290655.92269516 1.366921306 54.98662186 1.373521209

Example 5: Mild Sulfonation of Poly(Divinylbenzene) Based UncoatedPorous Polymeric Beads with Acetyl Sulfate, Followed byFunctionalization with Poly(N-Vinylpyrrolidone) as a HemocompatibleCoating, Used to Prepare Modified Polymer CY15048

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Among various “mild” sulfonating agents acetyl sulfate (prepared from98% conc. Sulfuric acid and acetic anhydride at low temperatures) isknown to be very efficient for DVB or styrene based polymeric materials.Sulfonation is usually done at 50° C. for several hours using equimolaramounts of acetyl sulfate and DVB or styrene based polymers. Sulfonationoccurs mainly at benzene ring and unreacted double bonds (in DVB basedcross-linked polymeric porous beads) could be preserved for furtherfunctionalization. Usually after sulfonation with acetyl sulfate, thepolymer is converted into ˜SO₃Na form and can be graft copolymerizedwith N-vinyl pyrrolidone (in bulk with benzoyl peroxide as initiator) orin water solutions (using sodium persulfate initiator). Resultingsulfonated polymer is “coated” with poly(N-vinylpyrrolidone) to makehemocompatible material capable of removing K⁺ cations fromphysiological fluids.

The base polymer selected for this modification was polymer CY15044. Thesulfonation and workup were carried out as described in Example 4, using45.0 g dry CY15044 polymer, 150 mL glacial acetic acid, 62.0 g aceticanhydride, and 40.0 g concentrated sulfuric acid. The resultingsulfonated polymer, in ˜SO₃Na form, was rewet in DI water in a 1 Ljacketed reaction vessel fitted with a Teflon coated agitator. DI waterwas removed from the vessel, and a solution composed of 75 mL NVPmonomer, 1.7 g sodium persulfate, and 25 mL DI water was added. Thereaction was allowed to proceed for 72 hours at 70° C. with agitationspeed set to 100 RPM. Resulting poly(NVP) coated polymer was washed fivetimes using 200 mL DI water, and dried in a vacuum oven until no furtherloss on drying was observed. Cumulative pore volume data for polymerCY15048 is shown below, in Table XX. A log differential pore volume plotis shown in FIG. 7.

Thrombogenicity was measured by the uPTT assay in which materials werecompared to the negative control (plasma alone), positive control (glassbeads) and reference beads to determine the degree of contact activationactivity. In the uPTT assay, the % change in clot formation over time ascompared to the reference materials was determined, then groupedaccording to: <25% activators, 25-49% moderate activators, 50-74% mildactivators, 75-100% minimal and >100% non-activators of the intrinsiccoagulation pathway. Polymer CY15048, 94%, was a minimal activator.

TABLE XX Nitrogen Desorption Data for CY15048 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 4355.2-828.4 944.3942734 0.0001057 828.4-474.0 558.6159743 0.000283797 474.0-303.3351.2170892 0.000357942 303.3-224.3 251.4508466 0.000719278 224.3-216.6220.2880162 0.001144201 216.6-208.1 212.161881 0.001932015 208.1-143.6163.4823625 0.075370749 143.6-108.1 120.3913517 0.239861146 108.1-81.7 90.85045572 0.292261423 81.7-71.8 76.00202402 0.305608258 71.8-60.264.83123041 0.320783836 60.2-52.6 55.8248996 0.329250736 52.6-46.649.1740518 0.335209946 46.6-41.3 43.5462226 0.339923553 41.3-37.439.07691383 0.349323983 37.4-32.5 34.52204235 0.351977397 32.5-29.430.76412188 0.352966945 29.4-27.3 28.24001433 0.353787455 27.3-24.625.76447932 0.355166927 24.6-22.3 23.26898468 0.357207636 22.3-19.920.87422635 0.360962494 19.9-17.5 18.46778237 0.367188172

Example 6: Mild Sulfonation of Poly(Styrene-co-Divinylbenzene) UncoatedPorous Polymeric Beads, Followed by Functionalization withPoly(N-Vinylpyrrolidone) as a Hemocompatible Coating, Used to PrepareModified Polymer CY15049

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

The base polymer selected for this modification was polymer CY15042. Thesulfonation and workup were carried out as described in Example 4, using45.0 g dry CY15042 polymer, 200 mL glacial acetic acid, 62.0 g aceticanhydride, and 40.0 g concentrated sulfuric acid. The reaction wasallowed to proceed for 2 hours. The resulting dried sulfonated polymer,in ˜SO₃Na form, was added to a 1 L jacketed reaction vessel fitted witha Teflon coated agitator. 140.0 g N-vinylpyrrolidone monomer and 2.0 gbenzoyl peroxide were added to the reactor. The reaction was allowed toproceed for 24 hours at 70° C. with agitation speed set to 100 RPM.Resulting poly(N-vinylpyrrolidone) coated polymer was washed five timesusing 200 mL DI water, and dried in a vacuum oven until no further losson drying was observed. Table XXI, below, displays cumulative porevolume data for polymer CY15049. FIG. 8 presents a log differential porevolume plot.

TABLE XXI Nitrogen Desorption Data for CY15049 Pore Diameter AverageCumulative Pore Range (Å) Diameter (Å) Volume (cm³/g) 6798.1-997.4 1113.294549 0.002499046 997.4-529.0 628.6356118 0.005782394 529.0-503.0515.3445059 0.00652485 503.0-431.3 461.4274588 0.007796961 431.3-320.8359.3702778 0.010896953 320.8-317.4 319.0643487 0.011833304 317.4-274.0292.3669097 0.013396248 274.0-230.2 248.1049882 0.016483381 230.2-225.4227.7447013 0.017366617 225.4-211.6 218.0383103 0.018833905 211.6-195.5202.8978228 0.029306436 195.5-143.0 160.6741284 0.494786051 143.0-99.0 112.5005572 0.779812896 99.0-82.5 89.05063735 0.848450234 82.5-69.874.92200629 0.902458565 69.8-59.0 63.37885992 0.947842682 59.0-51.254.4553105 0.981969695 51.2-44.8 47.47101253 1.011973922 44.8-39.441.69146063 1.039279282 39.4-35.3 37.10279099 1.066468142 35.3-31.333.03019211 1.096075821 31.3-28.2 29.57468036 1.118921801 28.2-25.526.70738162 1.143080339 25.5-22.8 23.944141 1.17115692 22.8-20.421.43590284 1.201324419 20.4-18.2 19.13416412 1.23163662

Example 7: Single-Pass Filtration for Hemoglobin and Potassium Removal

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Units of human pRBC were allowed to equilibrate to room temperature for30 minutes, where the units were gently mixed for 15 minutes. A bloodspike was inserted into the unit and samples for the initial hemoglobin(Hb) and potassium concentrations were taken. The blood spike line wasattached to the top port of the polymer containing filtration device,and a sample collection line attached to the bottom port. A pinch clampwas fitted on the sample collection line for flow control. Approximatelyone bed volume, 30 mL, was flushed through the device into a wastecontainer to purge the device of normal saline solution. The samplecollection tube was placed over 15 mL conical tubes where 12 mLfractions of pRBCs were collected at a flow rate of about 3-3.5 mL/minuntil the unit was completely filtered. Sample tubes were centrifugedfor 15 minutes at 4600 RPM at 4° C. Plasma supernatant from each sampletube was collected and the plasma free hemoglobin level was determinedby an absorbance read at 450 nm and potassium levels were measured witha potassium ion-selective electrode. The percentage of initial freehemoglobin removed during single-pass filtration, averaged from threetrials, is presented in FIG. 9. FIG. 10 displays pre- andpost-filtration potassium ion concentration in blood, averaged fromthree trials. Polymers CY15101 and CY15102 are able to removesignificant quantities of both potassium and hemoglobin, while polymerCY15100 only removes the potassium and does not remove hemoglobin.

Example 8: Dynamic Recirculation Filtration

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Polymer CY14144 has been tested in a dynamic competitive systemevaluating albumin (30 mg/mL) and myoglobin (100 mg/L) removal from aPBS solution with 8 mEq/L potassium. This model has been designed toreflect clinical albumin and myoglobin (rhabdomyolysis) values. Thedynamic system allows for the continuous measurement of proteinadsorption by the polymer beads at two UV wavelengths. As long as thesurrogate proteins, such as albumin and myoglobin, have different UVabsorption profiles, the two protein surrogates can be measuredsimultaneously, providing competitive adsorption conditions. This allowsa rapid assessment of polymer performance for the simultaneousadsorption of target and non-target factors under flow conditions; a keyparameter to assess studies that balance sorption withhemocompatibility. The dynamic system has been fully calibrated(absorbance and flow conditions) and was used to measure binding with a6 mL polymer filled device at a flow rate of 6 mL/min for five hours atroom temperature. CY14144 has a robust myoglobin adsorption, potassiumremoval and demonstrated good selectivity with minimal albumin removal.Dynamic recirculation data for CY14144, averaged from 7 trials, is shownbelow in FIG. 11. The average potassium removal, measured as the percentreduction from initial quantity, was found to be 25.3% with a standarddeviation of 1.42.

Thrombogenicity was measured by the uPTT assay in which materials werecompared to the negative control (plasma alone), positive control (glassbeads) and reference beads to determine the degree of contact activationactivity. In the uPTT assay, the % change in clot formation over time ascompared to the reference materials was determined, then groupedaccording to: <25% activators, 25-49% moderate activators, 50-74% mildactivators, 75-100% minimal and >100% non-activators of the intrinsiccoagulation pathway. Shown below, in Table XXII, is a comparison ofthrombogenicity for two different potassium removing polymers. PolymerCY14144 exhibits minimal thrombogenic activity while still removingpotassium and myoglobin simultaneously in a dynamic recirculation modelin phosphate buffered saline (PBS). In comparison, potassium sorbentCY14022 is a moderate activator of the intrinsic coagulation pathway bythe uPTT assay and is ineffective in myoglobin removal.

TABLE XXII Myoglobin and Potassium Removal from PBS in a DynamicRecirculation Model Polymer uPTT Myoglobin Removal Potassium RemovalCY14144 87% 71.63% 25.3% CY14022 59% 5.94% 66.07%

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

Additionally, polymer CY14144 is able to remove significant levels ofpotassium from blood plasma in a dynamic recirculation model. The normalrange for blood potassium is 3.5-5 mEq/L while a patient suffering fromhyperkalemia might have blood potassium levels up to 7-7.5 mEq/L.Reperfusion of plasma with a starting concentration of potassium 7.45mEq/L through a device filled with polymer CY14144 under recirculationconditions that mimic the clinical application reduced the potassiumconcentration to 4.52 mEq/L (a 2.93 mEq/L reduction) in 5 hours.

The general description and the following detailed description areexemplary and explanatory only and are not restrictive of the invention,as defined in the appended claims. Other aspects of the presentinvention will be apparent to those skilled in the art in view of thedetailed description of the invention as provided herein.

1-27. (canceled)
 28. A biocompatible polymer system comprising at leastone polymer, said polymer comprising (i) a plurality of pores and (ii) asulfonic acid salt functionality; wherein the polymer's pore structurehas a total volume of pore sizes in the range of from 10 Å to 40,000 Ågreater than 0.1 cc/g and less than 5.0 cc/g dry polymer; and whereinthe polymer is in the form of a crosslinked polymer or a porous polymer;wherein the crosslinked polymer or porous polymer has been sulfonatedunder mild conditions that retain residual functionality of anyunreacted double bonds and chloromethyl groups.
 29. The biocompatiblepolymer system of claim 28 wherein the polymer is hemocompatible. 30.The biocompatible polymer system of claim 28 wherein an agent is used toimbue biocompatibility to the polymer system, said agent being either(i) heparin or (ii) a heparin mimicking polymer.
 31. The biocompatiblepolymer system of claim 28 wherein the polymer is formed andsubsequently made to be biocompatible.
 32. The biocompatibility imbuingmodification of claim 31 wherein an agent used to imbue biocompatibilityis either (i) heparin or (ii) a heparin mimicking polymer.
 33. Thebiocompatible polymer system of claim 28 wherein the polymer system hasthe form of a solid support comprising a bead, fiber, monolithic column,film, membrane, or semi-permeable membrane.
 34. The biocompatiblepolymer system of claim 33 wherein the solid support has a biocompatiblehydrogel coating.
 35. The biocompatible polymer system of claim 28,wherein said polymer system is capable of adsorbing (i) protein basedtoxins or inflammatory mediators having a molecular weight of from about0.5 kDa to about 1,000 kDa and (ii) positively charged ions.
 36. Thebiocompatible polymer system of claim 28, wherein said polymer system iscapable of adsorbing (i) protein based toxins or inflammatory mediatorshaving a molecular weight of from about 1 kDa to about 1,000 kDa and(ii) positively charged ions.
 37. The biocompatible polymer system ofclaim 35 wherein, the toxins and inflammatory mediators comprise of oneor more of cytokines, superantigens, monokines, chemokines, interferons,proteases, enzymes, peptides including bradykinin, soluble CD40 ligand,bioactive lipids, oxidized lipids, cell-free hemoglobin, cell-freemyoglobin, DAMPS, growth factors, glycoproteins, prions, toxins,bacterial and viral toxins, PAMPS, endotoxins, drugs, vasoactivesubstances, foreign antigens, antibodies, and positively charged ions.38. The biocompatible polymer system of claim 28 wherein said polymer ismade using suspension polymerization, emulsion polymerization, bulkpolymerization, or precipitation polymerization.
 39. The biocompatiblepolymer system of claim 28 wherein said polymer is a hyper-crosslinkedpolymer.
 40. The biocompatible polymer system of claim 28, wherein theunreacted double bonds or chloromethyl groups can be modified via freeradical or S_(N)2 type chemistry to attach one or more of biocompatibleand hemocompatible monomers, cross-linkers or low molecular weightoligomers.
 41. The biocompatible polymer system of claim 28 wherein theporous polymer comprises sulfonic acid groups or a salt thereof,sulfonyl chloride, or sulfonate ester groups.
 42. The biocompatiblepolymer system of claim 41, wherein the polymer comprising sulfonic acidgroups or a salt thereof, sulfonyl chloride, or sulfonate ester groupsis produced by graft copolymerization of (i) premade porous polymer thatcontains unreacted double bonds with (ii) polymerizable vinyl monomerscontaining sulfonic acid groups or a salt thereof to form a mixturecomprising hemocompatible vinyl monomers.
 43. The biocompatible polymersystem of claim 28 constructed from polymerizable vinyl monomerscontaining sulfonic acid groups or a salt thereof which arecopolymerized in the presence of cross-linker, hemocompatible monomer,monomer and suitable porogen to yield porous polymeric polymercontaining a sulfonic acid salt functionality.
 44. The biocompatiblepolymer system of claim 28, wherein the polymer is housed in a containersuitable to retain the polymer and for transfusion of whole blood,packed red blood cells, platelets, albumin, plasma or any combinationthereof.
 45. The biocompatible polymer system of claim 28, wherein thepolymer is in a device suitable to retain the polymer and beincorporated into an extracorporeal circuit.
 46. The biocompatiblepolymer system of claim 28, wherein the crosslinked polymer is ahyper-crosslinked polymer.
 47. The biocompatible polymer system of claim28, wherein the porous polymer is a macroreticular porous polymer. 48.The biocompatible polymer system of claim 28, wherein the biocompatiblepolymer system has improved biocompatibility by use of (i) a coatingselected from a group consisting of poly(hydroxyethyl methacrylate),poly(hydroxyethyl acrylate), poly(dimethylaminoethyl methacrylate),salts of poly(acrylic acid), salts of poly(methacrylic acid),poly(diethylaminoethyl methacrylate), poly(hydroxypropyl methacrylate),poly(hydroxypropyl acrylate), poly(N-vinylpyrrolidone), poly(vinylalcohol), heparin-mimicking polymers and mixtures thereof or (ii) acoating comprising heparin; and
 49. The biocompatible polymer system ofclaim 28, wherein the polymer comprises residues of one or more monomersselected from acrylonitrile, butyl acrylate, butyl methacrylate, cetylacrylate, cetyl methacrylate, divinylbenzene, ethyl acrylate, ethylmethacrylate, ethylstyrene, methyl acrylate, methyl methacrylate, octylacrylate, octyl methacrylate, styrene, vinylbenzyl alcohol,vinylformamide, vinylnaphthalene, and vinyltoluene.
 50. A method ofperfusion comprising passing a physiologic fluid once through or by wayof an extracorporeal circuit through a device comprising thebiocompatible polymer system of claim
 28. 51. A device for removingprotein based toxins, inflammatory mediators and positively charged ionsfrom physiologic fluid comprising the biocompatible polymer system ofclaim
 28. 52. The device of claim 51 wherein said toxins andinflammatory mediators have a molecular weight of from about 0.5 kDa toabout 1,000 kDa.
 53. The device of claim 51 wherein said toxins andinflammatory mediators have a molecular weight of from about 1 kDa toabout 1,000 kDa.